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Offlinetwiztidunreal
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Registered: 07/14/13
Posts: 60
Last seen: 6 years, 5 months
First time LC
    #18796879 - 09/03/13 10:10 PM (10 years, 5 months ago)

I got 2 syringes from a site, 1 KS orig, 1 Creeper.  After I knocked up my first set of cakes, I thought I would try a LC of each, just to see.
I used pint jars, each with a tablespoon of Corn syrup, made self healing port, sterilized, and injected 2cc from each syringe into the appropriate jar.  That was on 8/20.  Here is what each looks like now as of 9/3.

LC of Creeper


LC of KS


I see growth in the creeper, but all creeper BFC's turned out with green mold.  The KS jar shows nothing, but all of the BFC jars from that syringe are growing amazingly...

Whats the deal?


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OfflinePussyFart
Retired Cultivation Extrodinaire
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Registered: 04/08/12
Posts: 22,502
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Last seen: 1 month, 11 days
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Re: First time LC [Re: twiztidunreal]
    #18797300 - 09/03/13 11:39 PM (10 years, 5 months ago)

LCs from spores are sketchy...

Actually, any LC not made from a clean agar wedge is sketchy...

It is recommended to just forget about LCs all together and focus on agar and G2G transfers for the best results.

The whole LC fad is soo 1999....


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OfflineNobitte
Student


Registered: 02/20/08
Posts: 493
Loc: Biosphere
Last seen: 6 years, 4 months
Re: First time LC [Re: PussyFart]
    #18797641 - 09/04/13 01:07 AM (10 years, 5 months ago)

I would advise using a Grain LC technique;

Add an agar wedge to about 200mls of grain in a jar, let it colonise, add 200ml or so of distilled water to the jar in aseptic conditions(doesn't have to be, i just prefer to pour it straight in, instead of injecting it), shake/vortex the grain around in the water (break it up first by bashing the jar around a bit), aspirate with fresh needles and syringes.

edit: 16g sharp or drawing up needles are best IME, dont bother making large syringes(>20cc) unless you need to.

Its not my idea, i just cant be bothered finding who did it first. You could probably do the whole process to some extent without opening the jar after the initial wedge inoculation (i don't, too fiddly)

You can see contamination and colonization clearly, you can shake to speed up colonization, the inoculate is ready to go, this is pretty much how i make all of my LC and it works marvels (G2G, while successful for me in the past seemed a bit laborious and error prone).


Edited by Nobitte (09/04/13 01:38 AM)


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OfflineRogerRabbitM
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Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 5 days
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OG Cultivator
Re: First time LC (moved) [Re: twiztidunreal]
    #18805390 - 09/05/13 09:54 PM (10 years, 5 months ago)

This thread was moved from Advanced Mycology.

Reason:
Not an advanced mycology topic.


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OfflineMush4Brains
LOOL HACKED!!!

Registered: 07/31/13
Posts: 4,419
Last seen: 9 years, 3 months
Re: First time LC [Re: Nobitte]
    #18805429 - 09/05/13 10:02 PM (10 years, 5 months ago)

Quote:

Nobitte said:
I would advise using a Grain LC technique;

Add an agar wedge to about 200mls of grain in a jar, let it colonise, add 200ml or so of distilled water to the jar in aseptic conditions(doesn't have to be, i just prefer to pour it straight in, instead of injecting it), shake/vortex the grain around in the water (break it up first by bashing the jar around a bit), aspirate with fresh needles and syringes.

edit: 16g sharp or drawing up needles are best IME, dont bother making large syringes(>20cc) unless you need to.

Its not my idea, i just cant be bothered finding who did it first. You could probably do the whole process to some extent without opening the jar after the initial wedge inoculation (i don't, too fiddly)

You can see contamination and colonization clearly, you can shake to speed up colonization, the inoculate is ready to go, this is pretty much how i make all of my LC and it works marvels (G2G, while successful for me in the past seemed a bit laborious and error prone).




GLC is fine.  Using a cake for an LC isn't the best idea.


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OfflineNobitte
Student


Registered: 02/20/08
Posts: 493
Loc: Biosphere
Last seen: 6 years, 4 months
Re: First time LC [Re: Mush4Brains]
    #18805980 - 09/06/13 12:50 AM (10 years, 5 months ago)

Who said anything about cakes?


--------------------
First we must learn...

Then... WE CAN TEACH :weirdeyes:


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InvisibleGretchenmeister
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Registered: 07/23/05
Posts: 1,032
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Re: First time LC [Re: Nobitte]
    #18817599 - 09/09/13 01:28 AM (10 years, 5 months ago)

I still love and use LC all the time. I am not picky about the source really. 

If you used a vendors syringe and/or you have inoculated spawn jars or cakes that appear to be growing a healthy culture, and you are sure you prepared and sterilized your LC properly,  then you can assume your contamination source came during inoculation at some point.

Some have better luck with LC than others, but I use it successfully all the time.  If I am using a vendors syringe, I ALWAYS use the first CC for LC and Agar cultures.  I would hesitate to use a syringe to make LC after already have knocked up jars with it. The risk of contams is just too great at that point. 

Typically you can use a quarter cc per pint (that's 4 lc's with one cc), or even 1/8th cc per half pint, which is what I prefer.  The reason I prefer the smaller jars is that with 1 cc I can make 8 jars of LC, and an extra drop to a couple plates.  This pretty much ensures, that with the first CC of my Vendors syringe, I will have clean live culture to start with within ten days, and also have a lasting culture to begin further isolation/propagation. 

Why 8 instead of 4? 1. Luck-hopefully beat contams by shear number of attempts. Assuming my culture was clean to start, if I personally fuck up a couple times....I still have several more excellent chances of success. 2. When I use LC I try not to go back to the LC jar more than 3ish times. The more you stick a jar the higher chances of comtam. Half pint LC's typically hold 120 mls, at least 80ml of which is useable and able to be sucked up (IME). That is a shitton of jars worth of inoculation juice.

You mention you have a RTV port, but don't mention whether you have a filter covered port/hole for GE?


--------------------
What's wrong with folks?

Point your IRC client to irc.socialirc.com, port 6667, #cultivation and #shroomery for live chat with like minded hobbyists and connoisseurs.


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OfflinePussyFart
Retired Cultivation Extrodinaire
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Registered: 04/08/12
Posts: 22,502
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Re: First time LC [Re: Gretchenmeister]
    #18817617 - 09/09/13 01:39 AM (10 years, 5 months ago)

Quote:

Gretchenmeister said:
If you used a vendors syringe and/or you have inoculated spawn jars or cakes that appear to be growing a healthy culture, and you are sure you prepared and sterilized your LC properly,  then you can assume your contamination source came during inoculation at some point.



Not really....no spore print/syringe is ever 100% clean really, vendor or not.

And GLC is not always going to be clean either, because we cannot observe every grain...

Just sayin...A majority of failed LCs are either from spores or GLC....hardly any when made from an agar wedge...

There is not much that can go wrong when using a red hot needle to inoculate...so unless you did not flame the needle, or did something stupid like wipe it with alcohol after flaming, I would not really assume it was from the inoculation.


Edited by PussyFart (09/09/13 01:39 AM)


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InvisibleGretchenmeister
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Registered: 07/23/05
Posts: 1,032
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Re: First time LC [Re: PussyFart]
    #18817635 - 09/09/13 01:50 AM (10 years, 5 months ago)

Quote:

Notahacker420 said:
Quote:

Gretchenmeister said:
If you used a vendors syringe and/or you have inoculated spawn jars or cakes that appear to be growing a healthy culture, and you are sure you prepared and sterilized your LC properly,  then you can assume your contamination source came during inoculation at some point.



Not really....no spore print/syringe is ever 100% clean really, vendor or not.

And GLC is not always going to be clean either, because we cannot observe every grain...

Just sayin...A majority of failed LCs are either from spores or GLC....hardly any when made from an agar wedge...

There is not much that can go wrong when using a red hot needle to inoculate...so unless you did not flame the needle, or did something stupid like wipe it with alcohol after flaming, I would not really assume it was from the inoculation.





I have great results for years and years.  I generally expect fifty fifty success rate, usually end up with 99% or better. 

I would think it is very safe to assume, if you are trying to break down the process and where a potential problem may have occurred, that if you have several jars of media colonizing successfully from the same source, that the introduction of contaminants to the LC could very well have occurred during the inoculation- IF one is sure the LC was prepared properly. 

I would never use a print to start an LC. I use vendors syringes to start them all the time though.  I am not saying LC is fail proof. I am saying it is not out dated, I have too much success with it for it to be outdated. Straight up.

I use grain LC quite often as well, usually shooting sterile water in a jar, sucking it out, and nocing up malt LC and/or some more spawn jars.  Works like a charm all day long, so long as you start with a clean jar and observe sterile procedures :smile:


--------------------
What's wrong with folks?

Point your IRC client to irc.socialirc.com, port 6667, #cultivation and #shroomery for live chat with like minded hobbyists and connoisseurs.


Mush Porn


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OfflinePussyFart
Retired Cultivation Extrodinaire
Male


Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
Last seen: 1 month, 11 days
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Re: First time LC [Re: Gretchenmeister]
    #18817644 - 09/09/13 01:54 AM (10 years, 5 months ago)

Quote:

Gretchenmeister said:
I am not saying LC is fail proof. I am saying it is not out dated, I have too much success with it for it to be outdated. Straight up.



I hear ya....to each his own....

Quote:

Gretchenmeister said:
I use grain LC quite often as well, usually shooting sterile water in a jar, sucking it out, and nocing up malt LC and/or some more spawn jars.  Works like a charm all day long, so long as you start with a clean jar and observe sterile procedures :smile:



I get that, but you cannot know for sure that every grain in the middle of the jar, that you cannot see, is "clean".

That is my only issue with GLC.


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